1. Field of the Invention
The present invention relates to a method for detecting a target nucleotide sequence using a complementary nucleotide sequence.
2. Background Art
A number of methods for detecting a target deoxyribonucleic acid (DNA) using a DNA that is complementary to the DNA sequence (xe2x80x9ccomplementary DNAxe2x80x9d) are well known. A typical example is the Southern blotting method to identify a specified DNA. Plaque hybridization and colony hybridization used in DNA cloning are also well known technique. The target DNA has first to be separated into a single strand as these methods are based on the fact that a target DNA hybridizes specifically to its complementary DNA.
However, with the exception of the Southern blotting method in which it is immobilized on a membrane, a single-strand DNA becomes spherical and such spherical DNA can not hybridize with the complementary DNA. Furthermore, it is necessary to heat a double-stranded DNA in order to convert it to a single-strand DNA. However such treatments adversely affect the target DNA.
The inventors found that the sensitivity of detecting a target nucleotide sequence using a complementary nucleotide sequence is remarkably improved by converting the target nucleotide sequence into a partially double-stranded nucleotide sequence. The present invention is based on this finding.
An object of the present invention is to provide a method of detecting a target nucleotide sequence using a complementary nucleotide sequence that has an excellent sensitivity of detection.
Another object of the present invention is to provide a method of producing a partially double-stranded nucleotide sequence that is used for the method of detecting the target nucleotide sequence.
The method of detecting a target nucleotide sequence according to the present invention comprises the steps of converting a target nucleotide sequence into a partially double-stranded nucleotide sequence and detecting the partially double-stranded nucleotide sequence using a complementary nucleotide sequence to the target nucleotide sequence (xe2x80x9ccomplementary nucleotide sequencexe2x80x9d).